Recent advances in optical microscopy, in particular those based on light sheet imaging, have significantly advanced the field of long term in vivo imaging with minimal perturbation to the sample. This means that there is now an interest in studying the processes by which a body repairs damage, in particular during development. A genetically encoded protein, KillerRed, is now available that is phototoxic, killing the cells in which it is present when illuminated with light at 561nm. The presentation will report for the first time its use in zebrafish with localised cell ablation in a SPIM system.
We report on ablation within the living sample either through the use of the light sheet within the microscope, but more precisely using a beam introduced through the imaging arm of a SPIM microscope controlled by adaptive optics to ensure localization of the activation. The beam profile is optimized to target individual cells within the kidney of the fish and the loss of fluorescence from the KillerRed is used to quantify the damage. The integration of the adaptive optics and opto-genetic encoding and activation of the KillerRed will be demonstrated to have the ability to ablate single cells deep within a living zebrafish with 100% survival of the fish. The presentation will illustrate how significant advances in the life sciences can be made through multidisciplinary research with optical expertise.
John Girkin, Charlotte Buckley, Mariana Carvalho, Laura Young, Sebastien Rider, and John Mullins, "Active beam shaping to optimize in vivo opto-genetic cell ablation (Conference Presentation)," Proc. SPIE 10502, Adaptive Optics and Wavefront Control for Biological Systems IV, 1050208 (Presented at SPIE BiOS: January 27, 2018; Published: 15 March 2018); https://doi.org/10.1117/12.2288682.5752144915001.
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